Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1993-09-23
1995-11-14
Knode, Marian C.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435220, 43525231, 435832, 25217412, C12N 952, C12N 954, C12N 170, C11D 940
Patent
active
054665949
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of PCT/DK92/00105 filed Apr. 3, 1992, which is incorporated herein by reference.
TECHNICAL FIELD
This invention is in the field of detergent proteases derived from strains of alkalophilic Bacillus sp. More specifically, the invention is directed towards a novel alkaline protease derived from a strain of Bacillus sp. JP 395. Moreover, the invention is directed towards a process for the preparation of the protease, the use of the protease as detergent enzyme, and detergent compositions comprising the protease of the invention.
BACKGROUND ART
Detergent enzymes have been marketed for more than 20 years and are now well established as normal detergent ingredients in both powder and liquid detergents all over the world. With the trend towards lower temperature washing, detergent enzyme consumption has increased during late years. Enzymes used in washing formulations comprise proteases, lipases, amylases, cellulases, as well as other enzymes, or mixtures hereof. Commercially most important are proteases.
Detergent proteases have been developed by isolation of proteases found in nature followed by testing in detergent formulations. Most detergent proteases are obtained from members of the genus Bacillus. Currently new types of proteases enter the market, offering the possibility of giving a better cost/performance ratio at various specified conditions.
Examples of commercial protease products are ALCALASE.TM., ESPERASE.TM. and SAVINASE.TM., all supplied by Novo Nordisk A/S, Denmark. These and similar enzyme products from other commercial sources are active in detergent solutions, i.e. at pH values in the range of from 8 to 11 and in the presence of sequestering agents, surfactants and bleaching agents such as sodium borate. The ALCALASE.TM. protease is produced by strains of the species Bacillus ficheniformis. The ESPERASE.TM. and SAVINASE.TM. proteases are obtained by cultivation of strains of alkalophilic Bacilli.
SUMMARY OF THE INVENTION
According to the present invention novel detergent proteases are provided.
In its first aspect, the invention provides a protease having an apparent molecular weight of 30 kD, a pl above 9.5, pH optimum in the range of from pH 9 to 11 (at 25.degree. C.), temperature optimum in the range of from 40.degree. to 50.degree. C. (at pH 9.5), and immunochemical properties identical or partially identical to those of a protease derived from Bacillus sp. JP 395, NCIMB No. 40337. in a more specific aspect, the protease is obtainable from a strain of Bacillus sp. JP 395. In a yet more specific aspect, the protease is obtainable from Bacillus sp. JP 395, NCIMB No. 40337, or a mutant or a variant thereof.
In another aspect, the invention provides an isolated biologically pure culture of a strain of Bacillus sp. JP 395. In a more specific aspect, a strain of Bacillus sp. JP 395, NCIMB No. 40337, or a mutant or a variant thereof, is provided.
In a third aspect, the invention provides a process for the preparation of the protease, which process comprises cultivation of a protease producing strain of Bacillus sp. JP 395 in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme. In a more specific aspect, Bacillus sp. JP 395, NCIMB No. 40337, or a mutant or a variant thereof, is cultivated.
In a fourth aspect, the use of the enzyme as detergent enzyme is claimed. In a more specific aspect, the invention provides a detergent composition and a detergent additive comprising the protease.
BRIEF DESCRIPTION OF DRAWINGS
The present invention is further illustrated by reference to the accompanying drawings, in which:
FIG. 1 shows the relation between temperature and the proteolytic activity of an enzyme according to the invention (the enzyme preparation obtained according to Ex. 1). Both curves were obtained with casein as the substrate at pH 9.5. However, the bottom curve was obtained in the presence of 0.1% STTP (sodium tripolyphosphate).
F
REFERENCES:
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P. Gemeiner et al., Folia Microbiol., vol. 36, No. 3, pp. 283-293, 1991.
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Y. Takii et al., Appl. Microbiol. Biotechnol., vol. 34, pp. 57-62, 1990.
Aaslyng Dorrit A.
Christiansen Margrethe
Dambmann Claus
Outtrup Helle
Knode Marian C.
Lambiris Elias J.
Nove Nordisk A/S
Weber Jon P.
Zelson Steve T.
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